THE ANATOMICAL RECORD PART A 288A:26 –35 (2006)
Specializations of the Granular
Prefrontal Cortex of Primates:
Implications for Cognitive Processing
GUY N. ELSTON,1,2* RUTH BENAVIDES-PICCIONE,3
ALEJANDRA ELSTON,1 BENDAN ZIETSCH,1 JAVIER DEFELIPE,3
PAUL MANGER,2 VIVIEN CASAGRANDE,4 AND JON H. KAAS5
1
Vision, Touch and Hearing Research Centre, School of Biomedical Sciences and
Queensland Brain Institute, University of Queensland, Queensland, Australia
2
School of Anatomical Sciences, Faculty of Health Sciences, University of the
Witwatersrand, Johannesburg, South Africa
3
Instituto Cajal (CSIC), Madrid, Spain
4
Department of Cell and Developmental Biology, Vanderbilt University,
Nashville, Tennessee
5
Department of Psychology, Vanderbilt University, Nashville, Tennessee
ABSTRACT
The biological underpinnings of human intelligence remain enigmatic. There remains
the greatest confusion and controversy regarding mechanisms that enable humans to conceptualize, plan, and prioritize, and why they are set apart from other animals in their
cognitive abilities. Here we demonstrate that the basic neuronal building block of the cerebral
cortex, the pyramidal cell, is characterized by marked differences in structure among primate
species. Moreover, comparison of the complexity of neuron structure with the size of the
cortical area/region in which the cells are located revealed that trends in the granular
prefrontal cortex (gPFC) were dramatically different to those in visual cortex. More specifically, pyramidal cells in the gPFC of humans had a disproportionately high number of spines.
As neuron structure determines both its biophysical properties and connectivity, differences
in the complexity in dendritic structure observed here endow neurons with different computational abilities. Furthermore, cortical circuits composed of neurons with distinguishable
morphologies will likely be characterized by different functional capabilities. We propose that
1. circuitry in V1, V2, and gPFC within any given species differs in its functional capabilities
and 2. there are dramatic differences in the functional capabilities of gPFC circuitry in
different species, which are central to the different cognitive styles of primates. In particular,
the highly branched, spinous neurons in the human gPFC may be a key component of human
intelligence. © 2005 Wiley-Liss, Inc.
Key words: extrastriate; cortex; pyramidal cell; human; macaque; marmoset;
aotus; galago; guenon; baboon; cortical surface area
Microanatomical studies have shown that the structure
of the most ubiquitous neuron in cortex, the pyramidal
cell, varies markedly between different cortical areas (Fig.
1). Up to a 30-fold difference has been reported in the
number of dendritic spines (the major postsynaptic sites of
excitatory inputs) on neocortical pyramidal cells in the
primate cerebral cortex (Elston et al., 2001, 2005h). Because pyramidal cells comprise over 70% of all neurons in
cortex (DeFelipe and Fariñas, 1992), these dramatic differences in their structure are likely to influence not only
cellular function, but the computational ability of the circuits they form (for reviews, see Shepherd and Greer,
1988; Churchland and Sejnowski, 1992; Koch, 1999; Mel,
©
2005 WILEY-LISS, INC.
1999; Segev et al., 2001; Elston, 2003a, 2006; Chklovskii
et al., 2004; London and Häusser, 2005). However, the
*Correspondence to: Guy N. Elston, Vision, Touch and Hearing
Research Center, School of Biomedical Sciences, University of
Queensland, St. Lucia, Queensland, 4072, Australia. Fax: 61-733654522. E-mail: [email protected]
Received 9 August 2005; Accepted 9 August 2005
DOI 10.1002/ar.a.20278
Published online 8 December 2005 in Wiley InterScience
(www.interscience.wiley.com).
GRANULAR PREFRONTAL CORTEX OF PRIMATES
Fig. 1. Plots of spine densities along the basal dendrites of pyramidal
cells in (top) the primary visual area (V1), (middle) the second visual area
(V2), and (bottom) granular prefrontal cortex (gPFC) in the human, baboon, macaque monkey, vervet monkey (guenon), marmoset monkey,
owl monkey (Aotus), and the galago. Note the differences in the profiles
in the different cortical regions. In V1, the plots of spine density are
remarkably tightly grouped in all primate species; in V2, there is some
degree of separation, but relatively little compared with that seen in
gPFC. Pyramidal cells in gPFC, particularly those in larger brained primates, are characterized by notably higher spine density along their
27
basal dendrites than those in V1 and V2. As each spine in cortex receives
at least one asymmetric synapse (putative excitatory input), differences
in spine density likely reflect variation in the number of excitatory inputs
to these neurons. Data for human, macaque, marmoset, owl monkey,
and galago modified from previous studies (Elston et al., 2001; Elston,
2003c; Elston et al., 2005b, 2005e, 2005h, 2005i). Data along the x-axis
have been displaced to avoid cluttering of the trend lines for all species
(all data were sampled at 0, 10, 20, 30, 40, 50, 60, . . ., ␮m from the cell
body).
28
ELSTON ET AL.
Fig. 2. Graphs illustrating the ratio of cortical surface area of granular
prefrontal cortex/cortical surface area of the frontal lobe (expressed as
percentage) vs. total cortical surface area (in mm2) for the human,
chimpanzee, gibbon, mandrill, baboon, macaque monkey, long-tailed
monkey, capuchin monkey, marmoset monkey, black lemur, and dwarf
lemur. Linear regression analysis revealed a positive slope (y ⫽
0.0003x ⫹ 42.547; r2 ⫽ 0.879), suggesting that prefrontal cortex has
expanded disproportionately during the evolution of these species. If
gPFC had not expanded, the ratio of gPFC/frontal lobe would result in a
regression with slope ⫽ 0. Prefrontal cortex is defined as granular cortex
anterior to the central sulcus, as observed in Nissl preparations (Brodmann, 1913).
data reported in these studies were not related back to the
size of the cortical area or brain, making it difficult to
determine whether variation in their structure parallels
cortical expansion.
Interspecies differences in pyramidal cell structure may
parallel the relative degree of expansion of particular cortical areas, lobes, the cortical mantle, or the entire brain.
Alternatively, variation in pyramidal cell structure may
reflect species-specific specializations that occur irrespective of size. Establishing which of these two possibilities
has occurred is essential if we are to better understand the
evolution of cortical circuitry and thus specialized cortical
function in different species (Kaas, 2000, 2005; Kaas and
Preuss, 2003). This is particularly pertinent to the study
of granular prefrontal cortex (gPFC), which has expanded
considerably in humans (Fig. 2) and is thought to be
important for executive cortical functions such as comprehension, planning, and perception (Goldman-Rakic, 1996;
Fuster, 1997; Barbas, 2000; Rolls, 2000; Miller and Cohen,
2001).
All animals were perfused by the same investigator
(G.N.E.) using the same protocol. Macaque (Macaca fasicularis) gPFC was taken from a 10-year-old male and
occipital cortex was taken from 4.5-year-old males (Elston,
2000; Elston et al., 2005a). Marmoset (Callithrix jacchus)
gPFC was taken from an 18-month-old male and occipital
cortex was taken from 24- to 27-month-old males (Elston
et al., 1999a, 2001). Tissue from the baboon (Papio ursinus) and vervet monkey (Cercopithecus pygerythrus) was
obtained from wild-caught adult males of unknown age
(Elston et al., 2005b, 2005c, 2005d, 2005e, 2005f, 2005g).
That from the owl monkey (Aotus trivirgatus) was obtained from a 21-year-old animal (Elston, 2003c) and that
taken from galago (Otolemur garnetti) was sampled from a
4-year-old animal (Elston et al., 2005h). All cases were
sexually mature. When possible (human, baboon, vervet
monkey, owl monkey, and galago), all tissue was sampled
from a single (left) hemisphere to avoid potential hemispheric and interindividual differences in cell morphology.
All experiments were performed in accordance with the
relevant guidelines for the care and use of animals in each
country in which the experiments were performed (United
States, Australia, Spain, Japan, and South Africa).
Blocks of tissue were taken from occipital and frontal
pole, including the primary (V1) and second (V2) visual
areas and dorsolateral gPFC, postfixed overnight as tangential preparations, and sections (250 ␮m thick) cut with
the aid of a Vibratome. The sections were then prelabeled
with 4,6-diamidino-2-phenylindole (Sigma D9542, St.
Louis, MO) to allow visually guided injection with Lucifer
Yellow (8% in 0.1 M Tris buffer, pH 7.4) by continuous
negative current under fluorescence illumination. Sec-
MATERIALS AND METHODS
Human tissue (Homo sapiens) was obtained within 2 hr
of death from a 48-year-old male who was killed instantaneously as a result of a car accident. The tissue was
obtained in accordance with the guidelines for the use of
human tissue of the Spanish Institutional Bioethical Committee (Spanish Council for Scientific Research). Once
excised, the tissue was immersed in 4% paraformaldehyde
for 24 hr (Elston et al., 2001). The other species were
overdosed by lethal injection of sodium pentobarbitone
and perfused intracardially with 4% paraformaldehyde.
29
GRANULAR PREFRONTAL CORTEX OF PRIMATES
TABLE 1 Cortical surface area (CSA), basal dendritic field area (BDFA) and total number of spines (TNS) for
pyramidal cells in the primary (V1) and second (V2) visual areas and granular prefrontal cortex (gPFC)
Species
Cortex
Variable
Otolemur
Callithrix
Aotus
Cercopithecus
Macaca
Papio
Homo
V1
CSA
BDFA
TNS
CSA
BDFA
TNS
CSA
BDFA
TNS
343a
48362k
556k
65a
64741k
1216k
—
118002u
3579u
341b
30600l
699l
98r
51700l,t
1240l,t
148d
105022t
3983t
400c
34400m
773m
95c
50400m
1459m
—
104000v
—
2156d
44301n
795n
—
82506n
1776n
1625d
130986w
5152w
1866e-i
43570o,p
735o,p
1203e,s
43900t
1139t
1733d
133000t
8766t
2559d
65019q
1077q
—
101210q
1962q
2111d
143672w
5009w
2826j
—
—
1454j
86000t
2417t
39287d
135000t
15138t
V2
gPFC
a
Rosa et al. (1997a).
Fritsches and Rosa (1996).
c
Tootell et al. (1985).
d
Brodmann (1913).
e
Felleman and Van Essen (1991).
f
Florence and Kaas (1992).
g
Horton and Hocking (1996).
h
LeVay et al. (1985).
i
Van Essen et al. (1984).
j
DeYoe et al. (1996).
k
Elston et al. (2005i).
l
Elston et al. (1999a).
m
Elston (2003b).
n
Elston et al. (2005e).
o
Elston and Rosa (1997).
p
Elston and Rosa (1998).
q
Elston et al. (2005b).
r
Rosa et al. (1997b).
s
Distler et al. (1993).
t
Elston et al. (2001).
u
Elston et al. (2005h).
v
Elston (2003c).
w
Present results.
b
tions were then processed with an antibody to Lucifer
Yellow [1:400,000 in stock solution, consisting of 2% bovine serum albumin (Sigma A3425), 1% Triton X-100
(BDH 30632, Poole, U.K.), 5% sucrose in 0.1 mol/l phosphate buffer], followed by a biotinylated species-specific
secondary antibody (Amersham RPN 1004, Arlington
Heights, IL; 1:200 in stock solution). The secondary antibodies were tagged with a streptavidin biotin-horseradish
peroxidase complex (Amersham RPN1051; 1:200 in phosphate buffer) and DAB (3,3⬘-diaminobenzidine; Sigma D
8001) was used as the chromogen (Elston and Rosa, 1997).
All cell injections were performed by two of the investigators (G.N.E. and R.B.-P.) and were standardized in all
studies.
Cells were drawn in two dimensions with the aid of a
camera lucida coupled with a Zeiss Axioplan microscope.
Dendritic tree size was determined using NIH Image (Bethesda, MD) by drawing a convex hull joining the outermost distal tips of dendrites of each cell. Twenty horizontally projecting basal dendrites of different cells in each
cortical area/species were drawn at high power (100⫻ oil
immersion) to quantify spine densities. Spine density was
determined per 10 ␮m segment of dendrite as a function of
distance from the cell body to the distal tips of the dendrites. All the above analysis was performed according to
blind procedures. The total number of spines in the dendritic trees was calculated as the sum of the product of
average number of dendritic intersections per annul derived from Sholl analysis and the average spine density
for the corresponding region of dendrites (Elston, 2001).
Moderated multiple regression (Aiken and West, 1991)
was used to determine significant differences between the
slopes of linear regression lines. All statistical analyses
were performed using SPSS software (SPSS, Chicago, IL).
RESULTS
Here we performed a systematic quantitative study of
pyramidal cell structure in V1, V2, and the gPFC of human, baboon, macaque monkey, vervet monkey, owl monkey, marmoset monkey, and the galago. Specifically, we
studied the size, branching structure, and total number of
spines in the basal dendritic trees of cortical pyramidal
cells at the base of layer III in gPFC (n ⫽ 220) and
compared these data with those sampled from layer III
pyramidal cells in V1 and V2 (n ⫽ 232 and 282, respectively). These morphological parameters differed appreciably between cortical areas/regions and species (Table
1). For example, in the macaque monkey, cells in the gPFC
were ⬎ 11 times more spinous than those in visual cortex.
Similar comparisons in the vervet monkey revealed a sixfold difference, greater than that observed in New World
marmoset monkey. Moreover, cells in the gPFC of humans
were 70% more spinous than those in the next closest
species (macaque monkey), three times more spinous than
30
ELSTON ET AL.
those in the baboon and vervet monkey, and more than
four times more spinous than those in the galago.
As each of these cortical areas/regions occupies a different absolute size in the cerebrum of the various species, it
was natural to ask whether there may be some underlying
trends in the regional and species differences in pyramidal
cell structure related to neocortical expansion. According
to the data published by Brodmann (1913), there is a
265-fold difference in the absolute size of in the gPFC of
the species included in the present investigation, with the
smallest observed in the marmoset (148 mm2) and the
largest in the human (3,928 mm2). A 22-fold difference
was observed in V2 and an 8-fold difference in V1 (Table
1). Comparison of the structure of pyramidal cells with the
absolute size of the cortical area/region in which they are
located revealed some interesting trends. Comparison of
the size of the basal dendritic trees of pyramidal cells with
the cortical surface area in V1, V2, and gPFC revealed an
increase in the two variables in all cortical regions (Fig.
3A). Moreover, the slopes returned by regression analysis
were remarkably similar for all three cortical regions,
suggesting a common trend in primates for increasingly
larger cells in increasingly larger cortical areas/regions.
However, comparison of our estimates of the total number of spines in the basal dendritic tree of the “average”
neuron in each cortical area revealed different trends in
gPFC, V1, and V2 (Fig. 3B). Specifically, the slope of the
linear regression for the gPFC data was considerably
steeper than that for either V1 or V2, suggesting that the
increase in the number of spines found in the dendritic
trees of pyramidal cells in gPFC during cortical expansion
far exceeds that in visual cortex. To test whether this
effect could be attributed to the increasing size of the
dendritic trees of the neurons, we plotted the total number
of spines in the dendritic tree vs. tree size in all three
cortical regions (Fig. 3C). These plots revealed two important observations: there is a progressive increase in the
linear regression slopes from V1 to V2 and gPFC, and
there is an extraordinary amount of variance in the gPFC
data not present in either V1 or V2. These data suggest
that in the gPFC the number of spines in the dendritic
trees of pyramidal cells more closely reflects the absolute
size of this region, rather than the size of their dendritic
trees.
To test whether or not these differences were significant, we performed a moderated multiple-regression analysis. By testing the relationship of any two predictors (e.g.,
V1/V2/gPFC and cortical surface area) on the criterion
(e.g., number of spines), and by testing the product of both
predictors (interaction term), we revealed significant differences in pairwise comparisons between cortical areas.
A significant increase in prediction at the second test
revealed a statistical difference (P ⬍ 0.05) between the
slopes of regression lines of gPFC and V2 for comparisons
between the total number of spines in the dendritic trees
of pyramidal cells and the cortical surface area (Fig. 3B;
r2change ⫽ 0.112). Significance was approached (P ⫽ 0.057)
for the comparison between the total number of spines in
the dendritic trees of pyramidal cells and the cortical
surface area for gPFC and V1 (Fig. 3B; r2change ⫽ 0.065).
Thus, human gPFC not only is considerably larger than
that in other primate species (in absolute and relative
terms), it is composed of pyramidal cells with highly complex dendritic trees studded with a disproportionately
high number of spines (putative excitatory inputs).
DISCUSSION
In the present study, we confirm and extend previous
reports of regional and species specialization in the neocortical pyramidal cell phenotype (for reviews, see Elston, 2002,
2006; Jacobs and Scheibel, 2002). We found, in many instances, dramatic differences in pyramidal cell structure
among V1, V2, and the gPFC in each of the seven different
primate species examined. Moreover, we found remarkable
phenotypic variation in pyramidal cell structure among the
different species in V1, V2, and gPFC. Comparison of the size
of the pyramidal cells in V1, V2, or gPFC revealed trends for
progressively larger neurons in species in which each of
these cortical areas/region occupied a larger absolute cortical
surface area. Furthermore, the rates of increase in cell size
and the surface area of the cortical area/region were similar
in V1, V2, and gPFC. Interestingly, however, we found different trends in the number of dendritic spines (putative
excitatory inputs) in the dendritic trees of pyramidal cells in
V1, V2, and gPFC of the different species. More specifically,
pyramidal cells in species with a relatively large gPFC were
disproportionately more spinous than those in species with a
relatively small gPFC (cells in human gPFC were 70% more
spinous than the next closest species, the macaque monkey,
and three times more spinous than those in the baboon and
vervet monkey). Moreover, the data reveal surprising variation in the pyramidal cell phenotype in the gPFC in primates not present in the occipital lobe. Thus, it appears as
though the recent and dramatic expansion of the gPFC in
primates has occurred not just by the addition of more neurons, but by the addition of more complex neurons. The
evolutionary and developmental mechanisms that influence
the complexity of the pyramidal cell phenotype remain to be
determined, as do their genetic and epigenetic regulation (for
reviews, see Nieuwenhuys, 1994; Marin-Padilla, 2001;
Preuss et al., 2004; Elston, 2006).
Structure/Function Relationship
The present data reveal that the extent to which different aspects of pyramidal cell structure (size and number of
spines) vary in different species depends on the cortical
region studied. Of particular interest here is that the
trend observed for the plots of the total number of spines
in the dendritic trees vs. either cortical surface area or size
of the basal dendritic trees in gPFC differs from that in V1
and V2 (Fig. 3B and C). How then might differences in the
size or spine density of pyramidal cells influence their
functional capabilities? While it is well known that neuron
structure determines its biophysical properties (Koch,
1999), it is less well known how neuron structure may
influence the functional properties of the circuits they
compose. It is our contention that cortical circuits composed of pyramidal cells of different structure will be characterized by different functional capabilities, much in the
same way that artificial systems composed of highly interconnected and powerful processors differ in their functional abilities to those composed of less powerful processors with fewer connections (for review, see Elston,
2003a). Several direct examples of the structure-function
relationship have been demonstrated between pyramidal
cell structure and cortical function. For example, differences in the size of the dendritic trees of pyramidal cells
potentially influence topographic sampling strategies and
mixing of inputs (Jacobs and Scheibel, 2002; Elston,
2003a). Differences in the branching complexity in the
GRANULAR PREFRONTAL CORTEX OF PRIMATES
Fig. 3. Plots of the (A) size of the basal dendritic trees of pyramidal
cells in the granular prefrontal cortex (gPFC), primary visual area (V1),
and second visual area (V2) vs. the total cortical surface area, (B) the
number of spines in the basal dendritic trees of pyramidal cells in V1, V2,
and PFC vs. the total cortical surface area, and (C) the number of spines
31
in the basal dendritic trees of pyramidal cells in V1, V2, and gPFC vs. the
size of the dendritic trees in human, baboon, macaque monkey, vervet
monkey (guenon), marmoset monkey, owl monkey (Aotus), and the
galago. Results of statistical comparisons are illustrated directly on the
plots. Asterisk, P ⬍ 0.05; number sign, P ⬍ 0.1.
32
ELSTON ET AL.
dendritic trees of pyramidal cells allow different degrees of
compartmentalization of processing of inputs (Poirazi and
Mel, 2001). Differences in the number of spines, each of
which receives at least one asymmetrical synapse, in the
dendritic trees of pyramidal cells reflect different numbers
of excitatory inputs sampled by individual cells (Harris
and Karter, 1994; Elston and DeFelipe, 2002). Thus,
highly branched and spinous pyramidal cells such as those
observed in human gPFC (Fig. 3B; see also Elston and
Zietsch, 2006) receive more putative excitatory inputs and
compartmentalize the processing of these inputs within
their dendritic trees to a great extent than smaller, less
spinous cells such as those in V1 and V2. These differences
in neuronal structure influence their potential for plastic
change (Stepanyants et al., 2002) and memory capacity
(Chklovskii et al., 2004), both thought to be important for
higher cortical functions.
Does this necessarily mean that more spinous cells such
as those in the gPFC of humans receive inputs from a
more diverse source of inputs than less spinous cells such
as those in the gPFC of galagos? Reciprocity dictates that
highly spinous neurons will receive more inputs than less
spinous neurons, providing a basis for increased recurrent
excitation (Wang, 2001; Jacobs and Scheibel, 2002; Elston,
2003a). However, very little is known of the diversity of
these inputs. While studies of connectivity in the cerebral
cortex are becoming increasingly more detailed (Hilgetag
et al., 1996; Melchitzky et al., 2001; Collins et al., 2005;
Germuska et al., 2005; Tanigawa et al., 2005), there exist
relatively few standardized comparative data. To the best
of our knowledge, only two such studies in gPFC have
been published (Bugbee and Goldman-Rakic, 1983; Preuss
and Goldman-Rakic, 1991). These authors studied patterns of connectivity in the gPFC of the New World squirrel monkey and the prosimian galago and compared them
with those in the gPFC of the macaque monkey. Notable
differences were detailed in the patterns of connectivity in
the gPFC of the macaque and both other species. However,
these studies need to be extended to include a greater
diversity of species, and the patterns of connectivity quantified. A systematic comparative study of patterns of connections to the gPFC of different primate species will no
doubt reveal new insights fundamental to a better understanding of cognition in primates.
There are, however, extensive data on patterns of connectivity in different regions of the cerebral cortex of the
macaque monkey, which reveal regional differences in the
diversity of inputs (for reviews, see Felleman and Van
Essen, 1991; Young, 1993). Comparison of these data with
those on regional differences in pyramidal cell structure
(Lund et al., 1993; Elston et al., 1999b, 2005a; Elston,
2000; Elston and Rockland, 2002) reveals some interesting
functional parallels. For example, highly branched, spinous neurons in gPFC are characterized by their tonic
activity, which is sustained despite intervention from distractors; less branched and less spinous neurons in sensory association cortex are characterized by tonic activity,
which desists following presentation of distractors; and
the least branched and least spinous cells in V1, for example, are characterized by phasic discharge properties
(Fuster and Alexander, 1971; Ashford and Fuster, 1985;
Koch and Fuster, 1989; Constantinidis and Steinmetz,
1996; Miller et al., 1996; Leung et al., 2002; Sakai et al.,
2002). Accepting the parallel between neuron structure
and function, the present results suggest that species dif-
ferences in prefrontal functions such as conceptual thinking, prioritizing, and planning may be attributed in part
at least to specializations in cortical microcircuitry. It
should be relatively easy to test the predictions in vivo.
For example, based on our data, we would expect that the
gPFC of the galago, for example, would be less adept at
sustaining tonic activity during presentation of distractors than that in the macaque monkey.
Methodological Considerations
Because of the difficulty in obtaining suitable material
from diverse primate species for comparisons (Crick and
Jones, 1993), selection of species included for study here has
been somewhat fortuitous. In order to be suitable for the cell
injection technique, tissue has to be obtained following surgical resection, perfusion, or postmortem. While tissue collected following surgical resection or perfusion may yield
large numbers of cells (⬎ 1,000 cells per case), tissue obtained postmortem is particularly problematic. Moreover,
sampling of diverse primate species is plagued with logistical
challenges; species are located in different geographical locations providing additional bureaucratic and political challenges. These experiments from which the present data are
drawn were performed over a period of 10 years in five
countries across four continents. Cell injection laboratories
were set up in each country, and the cell injection methodology was standardized in all studies. Species selection was
guided principally by the species Brodmann (1913) studied,
for which there exists a consistent set of quantified data (for
a translation, see Elston and Garey, 2004; see also Garey,
1994). Unfortunately, there are some glaring omissions of
species in the present study, including the gorilla, chimpanzee, and the orangutan, data from which would provide
crucial insights into evolutionary trends in pyramidal cell
structure.
Given these limitations on the selection of cases included here for analyses, it is natural to ask whether the
present results may be attributable to some form of sampling error such as interindividual variation or aging.
While we cannot rule out the possibility that these sources
of error may have influenced the results, it is unlikely that
they could account fully for our observations. For example,
in five of the seven species we injected cells in V1, V2, and
gPFC in a single hemisphere, discounting the possibility
of interindividual variation. Other experiments that show
consistent differences in pyramidal cell structure in somatosensory, motor, and cingulate cortex in different
cases make it increasingly less likely that the data presented here can be attributed to interindividual variation
(Elston et al., 2005c, 2005d, 2005f, 2005g). Based on our
previous cell injection studies in which we have sampled
neurons from a given cortical area in up to five different
individual age-matched animals, we would expect an interindividual variance in the total number of spines on
pyramidal cells of no more than 30% (e.g., Elston et al.,
2005a). To account for the differences reported here, interindividual variance in gPFC would have to be 22-fold
greater than that in V1 and 9-fold greater than that in V2
(i.e., a variance of 521 spines in V1, 1,278 spines in V2,
and 11,559 spines in gPFC).
Recently, Jacobs and colleagues revealed systematic differences in the maturation rate of pyramidal cells in different cortical areas (Travis et al., 2005), confirming and
extending their initial observations in V2 and gPFC (Jacobs et al., 1997). Moreover, dendritic and spine loss is a
GRANULAR PREFRONTAL CORTEX OF PRIMATES
common occurrence in aging (Nakamura et al., 1985;
Anderson and Rutledge, 1996; Duan et al., 2003). As the
data presented here were sampled from animals of different age, it is logical to question whether the results can be
attributed to age-related bias. While data have been published on the longevity of many primate species (Nowak,
1999), less is known of their relative developmental ages.
Here we restricted our analyses to mature animals, as
judged by their ages (when known), and the presence of
mature secondary sexual characteristics. As the gPFC
matures considerably later than visual cortex, any such
error would most likely result in underestimates in the
extent of differences reported between these two regions.
While estimates of the cortical surface area have been
calculated for individual cortical areas V1 and V2, the
estimates of the size of the gPFC include many cortical
areas (Brodmann, 1909). The relative size and number of
cortical areas within the gPFC may vary between species
(Brodmann, 1909). While quantitative comparative data
exist for the gPFC of humans and the macaque monkey
(Petrides and Pandya, 2001), little or nothing has been
published on the other species included here. Thus, it is
possible that some of the variance in the gPFC data may
be attributed to species differences in the size of the cortical areas in the gPFC. A more accurate depiction of the
trends illustrated in the gPFC of different primate species
is contingent on these data becoming available.
Because of the size of the dendritic trees of some pyramidal cells (up to 1 mm in diameter), it is difficult to inject
large numbers of cells in transverse cortical slices and
visualize their entire dendritic trees. To do so would require that cells be injected in cortical slices ⬎ 1 mm thick.
Normal biological variability in the trajection of the apical
dendrite, and asymmetries in the dendritic tree, would
require that sections ⬎ 1.5 mm thick be used, and cell
bodies located at a depth of 0.6 – 0.8 mm be injected, to
maximize the possibility of reconstructing the entire dendritic tree. However, it is not possible to visualize cell
bodies at such a depth sufficiently well to allow injection
under visual guidance, making it impossible to sample
sufficient numbers of cells in each case to allow meaningful statistical comparisons. Thus, we have focused our
study on the basal dendrites of pyramidal cells, as seen in
tangential sections. Moreover, we have restricted the
present analyses to layer III pyramidal cells. However,
our observations of the basal dendrites of infragranular
pyramidal cells suggest that they too differ markedly between cortical areas, and the nature of these differences
reflects those observed in supragranular pyramidal cells
(Elston and Rosa, 2000). Moreover, our observations of the
apical dendrites of pyramidal cells, although somewhat
limited, suggest that they too differ in their tangential
extent, branching complexity, and spine density in much
the same way as reported for the basal dendritic trees
(data not shown). However, further systematic comparative studies are required to provide more detail and a
better understanding of regional and species specializations in cortical circuitry.
Two other possible sources of error pertain to the determination of the cortical layer in which neurons were injected and the sampling strategy adopted for intracellular
injection. In the first instance, all cortical areas included
in the present study have a granular layer (layer IV). We
use the nomenclature of Hassler (1966) in preference to
that used by Brodmann (1909) for reasons outlined in
33
Casagrande and Kaas (1994) and Elston and Rosa (1997).
In V1, we sampled cells immediately above the granular
layer, corresponding to layer IIIC of Hassler and layer
IVC␣ of Brodmann. In V2 and gPFC, cells were sampled at
the base of layer III according to the nomenclature of both
Hassler and Brodmann. The granular layer is easily distinguished in DAPI-labeled sections by virtue of the high
cell density and small cell body size (see Fig. 2 of Elston
and Rosa, 1997). Thus, despite differences in the overall
thickness of cortex among cortical areas and species, we
were able to identify the section that contained the granular layer and select the adjacent section for intracellular
injection (that section closer to the cortical surface, which
contained layer III).
In the second instance, DAPI-labeled cells were injected
pseudorandomly. Cells are injected in a grid-like pattern
allowing even sampling across cortex. Thus, pyramidal
cells that project within the cortical area, to other cortical
areas (ipsi- and contralateral), and to subcortical targets
were all likely to have been included for analyses. Because
the structure of these cell types differs even within a
cortical area, some of the trends presented here may reflect variation in the proportion of these cell types among
cortical areas and species. This possibility does not detract
from our main findings, that is, there exist regional and
species differences in pyramidal cell structure, which are
likely to influence patterns of connectivity and function.
Interestingly, however, while the structure of these projection-identified pyramidal cells may be distinguishable
within a cortical area, the structure of each type differs
markedly between cortical areas (e.g., callosally projecting
or corticocortically projecting cells), leading to the conclusion that evolutionary and developmental mechanisms
that determine patterns of arealization and lamination in
cortex provide a stronger influence over pyramidal cell
structure than the targets to which they project (Vercelli
and Innocenti, 1993).
SUMMARY AND CONCLUSIONS
The granular prefrontal cortex has undergone dramatic
expansion in primates. This expansion has not occurred
simply by the addition of more neurons, but by the addition of more complex neurons. Specifically, there is a dramatic increase in the branching complexity and number of
spines in the dendritic trees of pyramidal cells in the
human gPFC. The disproportionately high number of
spines in the dendritic trees of pyramidal cells in the
human gPFC suggests they receive more excitatory inputs
than their counterparts in other primate species. The increase in neuron number, coupled with the increasing
potential to sample progressively more inputs, makes it
likely that the human gPFC is characterized by more
complex connectivity than that in other species. Species
differences in the gPFC pyramidal cell phenotype, and the
circuits they compose, are likely to influence functions
associated with this region such as comprehension, planning, and perception. In particular, species differences in
the gPFC pyramidal cell phenotype are likely to influence
their cognitive styles.
ACKNOWLEDGMENTS
The authors thank Jack Pettigrew for his comments on
a previous version of this manuscript. Supported by
grants from the J.S. McDonnell Foundation and the Na-
34
ELSTON ET AL.
tional Health and Medical Research Council of Australia
(to G.N.E.), the Ministry of Education and Science of
Spain (to J.d.F.), the Comunidad Autónoma de Madrid (to
R.B.-P.), and the South African National Research Foundation (to P.M.).
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